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. Author manuscript; available in PMC: 2015 Sep 4.
Published in final edited form as: Mol Cell. 2014 Nov 13;56(5):696–707. doi: 10.1016/j.molcel.2014.10.011

Figure 5. Wisp overexpression leads to miRNA adenylation and impairs target repression activity.

Figure 5

(A) Northern blot analysis following co-transfection of the miR-312 and Wisp expression plasmids into S2 cells. miR-312 is highly modified and decreased in cells overexpressing Wisp, but not affected by the catalytic mutant (DADA). This is a representative result of three independent experiments. (B) Small RNA sequencing of the following samples: “Mock” (S2 cell transfected with only miR-312) and “Wisp expression” (S2 cell transfected with miR-312 and Wisp). Reads of miR-312 aligned to the reference sequence. The RPM values of each sequence are presented on the left side. (C) Dual luciferase reporter assay to measure miR-312 activity. A firefly luciferase reporter containing two miR-312 bulged sites was used. The relative activity was calculated by firefly luciferase activity from the miR-312 reporter (normalized to renilla luciferase activity) divided by that of firefly luciferase activity from the mutant reporter (normalized to renilla luciferase activity). Data are presented as mean ± standard error of four biological replicates. The p value was determined by two-tailed T-test (*: p < 0.05, **: p < 0.01). See also Figure S3.